CN110025618A - Artesunate combines the application in the liver cancer Huh7 cell drug for preparing AKTc-Myc activation with Sorafenib - Google Patents
Artesunate combines the application in the liver cancer Huh7 cell drug for preparing AKTc-Myc activation with Sorafenib Download PDFInfo
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Abstract
The invention discloses a kind of Artesunates to combine the application in the drug of the human liver cancer Huh7 cell of preparation treatment AKT/c-Myc activation with Sorafenib.Artesunate combines the application in the drug of the liver cancer Huh7 cell of preparation treatment AKT/c-Myc activation with Sorafenib, the Artesunate can reduce the protein level of p-AKT, c-Myc, to inhibit liver cancer Huh7 cell Proliferation, the Artesunate combines the protein level by reducing p-AKT, c-Myc with Sorafenib, further more inhibits the growth of liver cancer Huh7 cell.Therefore, the drug candidate that Artesunate and Sorafenib combination can be used as anti-human liver cancer cell Proliferation is researched and developed.
Description
Technical field
The present invention relates to a kind of cancer treatment drugs, belong to biopharmaceutical technology, and in particular to Artesunate and rope
Application of the La Feini combination in the drug of the Huh7 cell of preparation treatment AKT/c-Myc activation.
Background technique
Hepatocellular carcinoma (Hepatocellular carcinoma, HCC) is one of most common malignant tumour of liver, is
The great cancer of death rate the second in the world.The concealment of liver cancer onset, it is unobvious without symptom or symptom in early days, it is not easy to be found, only
40% patient can be discovered in an early phase.Locally Advanced is had reached when Most patients are made a definite diagnosis or DISTANT METASTASES IN occurs, and treatment is tired
Difficulty, prognosis is very poor, seriously threatens health of people.
Artesunate is the antimalarial agent of China's classics, is a kind of derivative of qinghaosu, and English name is
Artesunate, molecular formula C19H28O8, molecular weight 384.42, chemical structural formula such as formula (I), colourless, bitter.Studies have shown that
Artesunate also has antitumor characteristic.
Sorafenib (Sorafenib) is the current clinical unique targeted drug for being directed to advanced liver cancer patient.Sorafenib
It is first is also uniquely by U.S. Food and Drug Administration and European drugs administration approved and to be used to treatment evening
The first-line drug of phase liver cancer.Two medicine use in conjunction are in terms of liver cancer treatment.Home and abroad report is less.Explore good effect, safety
Height, the systemic therapy scheme for being more suitable for advanced hepatocellular carcinoma patient have important Clinical significance of MG.Sorafenib English name
Referred to as sorafenib, molecular formula are as follows: C21H16ClF3N4O3, molecular weight 464.825, chemical structural formula such as formula (II).
Summary of the invention
Of the existing technology in order to solve the problems, such as, the purpose of the present invention is to provide a kind of Artesunate and Sorafenibs
The application in the Huh7 cell drug that AKT/c-Myc is activated is combined, is a kind of biological therapy tumor candidate drug Artesunate
It is combined with Sorafenib and is preparing the application in antitumor.
The present invention is achieved through the following technical solutions:
Artesunate combines answering in the drug of the Huh7 cell of preparation treatment AKT/c-Myc activation with Sorafenib
With the Artesunate can reduce the protein level of p-AKT, c-Myc, to inhibit liver cancer Huh7 cell Proliferation, the blueness
Artemisic succinate combines the protein level by reducing p-AKT, c-Myc with Sorafenib, further more inhibits liver cancer Huh7 cell
Growth.
Application of the present invention, the Artesunate are mentioned from compositae plant artemisia annua Artemisia annual
It takes, molecular formula C19H28O8, it is -10 α of dihydroartemisinine-succinate monoester, is calculated by anhydride, contains C19H28O8It should be
98.0%~102.0%, belong to white crystalline powder, odorless bitter is readily soluble in ethyl alcohol, acetone or methylene chloride, adds two
Chloromethanes makes to dissolve and quantify the solution for diluting and being made in every 1ml containing 25mg, measures in accordance with the law, and specific rotation is+4.5 ° to+6.5 °,
PH value is 3.5~4.5, and preparing fusing point is 1290 DEG C~1400 DEG C, is now widely used for treatment malaria, and toxicity is smaller.
Application of the present invention, the Sorafenib are also known as Raf inhibitor, and in December, 2005 is through food and medicine office, the U.S.
(FDA) ratify the first-line drug listing as treatment tumour.For white to gray solid, molecular formula C21H16ClF3N4O3, heat
It has good stability, does not absorb water, be slightly soluble in ethyl alcohol, be dissolved in polyethylene glycols 400, fusing point is 202 DEG C -204 DEG C.
Application of the present invention, the Artesunate are combined with Sorafenib, concentration >=12.5 μ of Artesunate
mol·L-1, the concentration >=2.5 μm olL of Sorafenib-1, the composition of medicine can be made into tablet, preparation method are as follows:
1) Artesunate 12.5g, Sorafenib 2.5g are weighed, it is mixed that grinding in mortar is added in starch 8.0g, dextrin 12.0g
It is even；
2) tartaric acid of 0.4g is dissolved in 50% ethyl alcohol, is repeatedly added in mixed-powder by appropriate amount, when addition disperses
Face is big, is uniformly mixed, softwood is made；
3) wet grain is made by 18~20 mesh nylon mesh, 60 DEG C or less dryings can rise to 70 DEG C hereinafter, accelerating dry when close dry
Dry, dry granular moisture control is below 1.5%；
4) go out whole grain by 20 meshes, mix with the magnesium stearate of 0.4g sieving, then mixed again with dry particl, calculating contains
Amount carries out punch die tabletting with 8mm punch die；Up to Artesunate Sorafenib piece.
Application of the present invention, the composition of medicine can be made into freeze drying powder injection, preparation method are as follows:
1) it is swollen HYDROXYPROPYL BETA-CYCLODEXTRIN: weighing 180g HYDROXYPROPYL BETA-CYCLODEXTRIN, be placed in 1L beaker, be added
400ml water for injection, magnetic agitation (revolving speed 500rpm) 3h in 50 DEG C of water-baths, swelling are spare.
2) it prepares inclusion compound: weighing 2.5g Sorafenib and 12.5g Artesunate, be placed in 2L beaker, addition is swollen
HYDROXYPROPYL BETA-CYCLODEXTRIN solution, 800rpm stir 10min until dissolution.
3) it adjusts pH and 72g mannitol is added, continue stirring to after dissolving, arrived with the pH value of 1mol/L hydrochloric acid conditioning solution
10.1, solution is settled to 900ml with water for injection.
4) 900mg active carbon is added in depyrogenation, after 500rpm magnetic agitation adsorbs 30min, 0.22 μm of filtering with microporous membrane.
5) filling, freeze-drying rolls lid to get Artesunate Sorafenib freeze drying powder injection.
The invention has the advantages that and advantageous effects:
1) present invention discover that Artesunate is inhibited to human liver cancer Huh7 cell, and good time and dense is presented
Dependence is spent, cell viability experimental result is shown, it is higher in the Artesunate concentration of same action time, the proliferation of cell is pressed down
Rate processed is higher；Cell clonal formation experimental result is shown:
Artesunate significantly inhibits human liver cancer Huh7 cell Proliferation, when Artesunate concentration is 6.25 μM
When, the Colony formings of liver cancer cells inhibit 50% hereinafter, and as Artesunate concentration increases, inhibitory effect is better.
2) present invention discover that Artesunate can be by reducing the expression of P-AKT, c-Myc, and then inhibit human liver cancer Huh7
Cell Proliferation.
3) present invention discover that Artesunate is combined with Sorafenib can more inhibit human liver cancer Huh7 cell Proliferation, cell
Colony formation is as the result is shown: when 12.5 μM of Artesunate, 2.5 μM of Sorafenib, the Colony forming of liver cancer cells inhibits
50% hereinafter, and with the increase of Sorafenib concentration, inhibitory effect is better.
4) present invention discover that Artesunate combined with Sorafenib can by reduce P-AKT, c-Myc protein expression,
And then inhibit human liver cancer Huh7 cell Proliferation, increase the susceptibility of Sorafenib.
5) present invention provides scientific basis to develop new antitumor drug candidate, has to exploitation new Chinese medicine important
Meaning.
Detailed description of the invention
Specific embodiments of the present invention will be described in further detail with reference to the accompanying drawing, in which:
Fig. 1 is that cell viability tests the Artesunate for detecting various concentration under different time to human liver cancer in embodiment 1
The inhibited proliferation figure of Huh7 cell；
Fig. 2 is the Artesunate of cell clonal formation experiment detection various concentration in embodiment 1 to human liver cancer Huh7 cell
Inhibited proliferation figure；
Fig. 3 is the Artesunate of cell clonal formation experiment detection various concentration in embodiment 1 to human liver cancer Huh7 cell
Inhibited proliferation quantitatively scheme；
Fig. 4 is that Western blot detects Artesunate to the influence diagram of AKT/c-Myc protein level in embodiment 2.
Fig. 5 is that influence of the Western blot detection Artesunate to AKT/c-Myc protein level is quantitative in embodiment 2
Figure.
Fig. 6 is that the Sorafenib of cell viability experiment detection various concentration in embodiment 3 is combined with Artesunate to people liver
The inhibited proliferation figure of cancer Huh7 cell.
Fig. 7 is that cell clonal formation experiment detects Artesunate and Sorafenib pharmaceutical composition to human liver cancer in embodiment 3
The inhibited proliferation figure of Huh7 cell.
Fig. 8 is that cell clonal formation experiment detects Artesunate and Sorafenib pharmaceutical composition to human liver cancer in embodiment 3
The inhibited proliferation of Huh7 cell is quantitatively schemed.
Fig. 9 is that Western blot detection Artesunate is combined with Sorafenib to AKT/c-Myc albumen table in embodiment 4
Up to horizontal influence diagram.
Figure 10 is that Western blot detection Artesunate is combined with Sorafenib to AKT/c-Myc albumen in embodiment 4
The influence of expression is quantitatively schemed.
Specific embodiment
Below with reference to specific implementation column and the chart technical solution that the present invention will be described in detail, but the present invention be not limited in
Under embodiment.
Artesunate is widely used for treatment malaria now, has extensive bio-pharmacology activity, wherein antitumor work
Concern of the property by many researchers.Sorafenib is unique targeted drug for the treatment of of late stage liver cancer, but due to congenital acquired
Drug resistance, extension patient vitals are limited, and the mechanism that Artesunate plays specific anticancer activity is still unclear, this may be with Sorafenib
It is combined into new anti-liver cancer and anti-drug candidate.The present invention explores Artesunate to the inhibiting effect of liver cancer cells Huh7, and hair
Existing antihepatocarcinoma effect mechanism provides basis to increase Sorafenib liver-cancer medicine susceptibility, is the treatment and diagnosis of liver cancer patient
New method is provided, is application based theoretical of the Artesunate in clinical liver cancer treatment.
The present invention provide a kind of biological therapy tumor candidate drug Artesunate combined with Sorafenib prepare it is antitumor
In application.The protein level for reducing p-AKT, c-Myc is combined with Sorafenib more particularly to Artesunate, to inhibit liver cancer
Huh7 cell Proliferation.
The present invention is tested by cell viability, cell clonal formation is tested, Western blot experimental study Artesunate
It is combined with Sorafenib to human liver cancer Huh7 inhibiting effect.
Various test apparatuses and reagent are commercial goods, are that can buy to obtain by commercial sources.Wherein, sweet wormwood amber
Ester purchase is 98% in sigma company, the U.S., product type A3731, Artesunate content, and Sorafenib is bought in the U.S.
Sigma company, product type A7397, Sorafenib content are 98%, and cell strain used in experiment in vitro is that source of people liver cancer is thin
Born of the same parents (Huh7) derive from U.S. ATCC cell bank.
Embodiment 1
Tablet, preparation method is made in the drug that Artesunate is combined with Sorafenib are as follows:
1) Artesunate 12.5g, Sorafenib 2.5g are weighed, it is mixed that grinding in mortar is added in starch 8.0g, dextrin 12.0g
It is even.
2) tartaric acid of 0.4g is dissolved in 50% ethyl alcohol, is repeatedly added in mixed-powder by appropriate amount, when addition disperses
Face is big, is uniformly mixed, softwood is made.
3) wet grain is made by 18~20 mesh nylon mesh, 60 DEG C or less dryings can rise to 70 DEG C hereinafter, accelerating dry when close dry
Dry, dry granular moisture control is below 1.5%.
4) go out whole grain by 20 meshes, mix with the magnesium stearate of 0.4g sieving, then mixed again with dry particl, calculating contains
Amount carries out punch die tabletting with 8mm punch die；Up to Artesunate Sorafenib piece.
Administration mode:
Adult usual amounts: oral administration, one time 4~6,2 times a day.
Points for attention: First Trimester is used with caution.
Embodiment 2
Freeze drying powder injection, preparation method is made in the drug that Artesunate is combined with Sorafenib are as follows:
1) swelling HYDROXYPROPYL BETA-CYCLODEXTRIN weighs 180g HYDROXYPROPYL BETA-CYCLODEXTRIN, is placed in 1L beaker, and 400ml is added
Water for injection, magnetic agitation (revolving speed 500rpm) 3h in 50 DEG C of water-baths, swelling are spare；
2) it prepares that inclusion compound weighs 2.5g Sorafenib and 12.5g Artesunate is placed in 2L beaker, is added and has been swollen
HYDROXYPROPYL BETA-CYCLODEXTRIN solution, 800rpm stirs 10min until dissolution；
3) it adjusts pH and 72g mannitol is added, continue stirring to after dissolving, arrived with the pH value of 1mol/L hydrochloric acid conditioning solution
10.1, solution is settled to 900ml with water for injection；
4) 900mg active carbon is added in depyrogenation, after 500rpm magnetic agitation adsorbs 30min, 0.22 μm of filtering with microporous membrane；
5) filling, freeze-drying rolls lid to get Artesunate Sorafenib freeze drying powder injection；
Medication: 100mg is injected in intravenous injection daily, continuous injection 3 days.
Points for attention: First Trimester is used with caution.
Effete test embodiment 1
The Artesunate of detection various concentration is under different time to the inhibited proliferation of human liver cancer Huh7 cell.
(1) Cell Viability Assay method measures cell Proliferation
Huh7 cell is taken out from incubator, digests, is centrifuged, count, with 3000 cell inoculations in 96 orifice plates, every hole
100 μ l culture mediums.After culture for 24 hours, corresponding drug is added, Artesunate concentration is respectively 0,12.5,25,50,100 μm of ol/
L, every group sets 5 multiple holes, continue culture for 24 hours, after 48h, 72h, the corresponding time take out, be added active agent CellTiter-Luminescent Assay carries out Luminescent cytoactive detection with microplate reader.Calculate the survival rate of cell.
Survival rate=A of cellDosing group/ABlank control group；
Cell viability experimental result is shown: as seen from Figure 1, the Artesunate (0,12.5,25,50,100 μM) of various concentration
Human liver cancer Huh7 cell strain is handled, CellTiter- is usedLuminescent Cell ViabilityAssay has detected blueness
Artemisic succinate act on for 24 hours, cell survival rate after 48h, 72h.The result shows that Artesunate is in human liver cancer Huh7 cell strain survival rate
Time-and concentration-dependent.
(2) cell clonal formation is tested
1) when cell grows into logarithmic phase, after being digested with pancreatin, the quantity of cell is measured with cell counter；
2) prepare clean 6 orifice plates, every hole number of plated cells are 5000；
3) after culture for 24 hours, artesunate drug, 0,12.5,25,50,100 μm of olL of concentration is added-1, 37 DEG C,
5%CO2Incubator in cultivate one week (7 days), take out and use violet staining, every hole is added 500 μ L Crystal Violet Dyes, is placed in and shakes
Bed is dyed for upper 30 minutes；
4) it is cleaned 3 times with PBS, five minutes every time, dries, take pictures after observation；
5) 10% glacial acetic acid of 1mL is added in every hole, is placed in 30 minutes on shaking table and decolourizes；
6) every hole takes 100 μ L to be placed in 96 orifice plates, and microplate reader measures the absorbance under 550nm；
7) data statistics: Cell colonies assay=ADosing group/ABlank group。
Cell clone is as the result is shown: by Fig. 2 and Fig. 3 as it can be seen that compared with blank control group, liver cancer cells Huh7 is by green
Artemisic succinate is handled one week, observes the formational situation of cell clone, and with the increase of concentration, clone's quantity of formation is reduced.?
12.5 μm of olL of Huh7 cell-1When cell obviously inhibit, only a small number of Clone formations.
Effete test embodiment 2
The variation that Western blot detection Artesunate expresses AKT/ signaling pathway protein.
The GAP-associated protein GAP of investigation has: P-AKT, AKT, c-Myc.
Huh7 cell is washed 3 times after the processing of the Artesunate of various concentration with the PBS of ice, and the RIPA that 200 μ l are added is thin
Cellular lysate liquid (contains protease inhibitors), with cell scraper by under cell scraper, collects into 1.5mlEP pipe, is inserted into ice and cracks
1h, during which overturn makes cell cracking abundant several times.Supernatant is taken to use after cell centrifugation (12000rpm, 20min, 4 DEG C) after cracking
BCA measures its protein concentration, mixes with sample-loading buffer, and 97 DEG C are boiled 10min, makes albuminous degeneration.
20 μ l, SDS- polyacrylamide constant pressure 100v electrophoretic separation albumen is sampled, then extremely by the protein delivery on gel
On nitrocellulose filter, 2h is closed with the TBST containing 5% skimmed milk power, is washed 4 times, each 10min, 4 DEG C of primary antibody overnight incubations,
TBST is added secondary antibody (1:3000-1:5000) and is incubated at room temperature 60min after washing 4 times, TBST is washed 4 times, and ECL luminescent solution is added and exposes
Light.Developer solution 1:1 matching while using, drop covers film, darkroom tabletting, automatic film developer development, fixing on film when use.
The above experiment is in triplicate.
Albumen is as the result is shown: such as Fig. 4 and Fig. 5 as it can be seen that compared with blank control group, after Artesunate is added, Huh7 is thin
The protein level of p-AKT, c-Myc intracellular is lowered, and difference has statistical significance (P < 0.05), to inhibit human liver cancer thin
Born of the same parents' Huh7 cell Proliferation.
Effete test embodiment 3
The Sorafenib of detection various concentration combines the inhibited proliferation to human liver cancer Huh7 cell with Artesunate.
(1) CellTiter-The detection of Luminescent Assay cell viability
Huh7 cell is taken out from incubator, digests, is centrifuged, count, with 3000 cell inoculations in 96 orifice plates, every hole
100 μ l culture mediums.After culture for 24 hours, corresponding drug is added, Sorafenib concentration is 0,0.5,1,2.5,5 μm of ol/L, sweet wormwood amber
Ester (12.5 μm of ol/L) is used in combination with Sorafenib (0,0.5,1,2.5,5 μm of ol/L), and every group sets 5 multiple holes, continues to cultivate
For 24 hours, it after 48h, 72h, is taken out in the corresponding time, active agent CellTiter- is addedLuminescent Assay,
Luminescent cytoactive detection is carried out with microplate reader.Calculate the survival rate of cell.
Cell survival rate=A drug/A control；
Cell viability is as the result is shown:
Using the Sorafenib of various concentration, concentration is (0,0.5,1,2.5,5 μM), according to the sweet wormwood of previous trial example 1
We have selected the concentration (12.5 μM) of Artesunate to the Activity Results of amber ester, after handling 48h, use
Luminescent Cell Viability Assay detect cell activity, individually handled with Sorafenib compared with, sweet wormwood amber
The activity of cell is inhibited by more conspicuousness when ester and Sorafenib are used in combination, and shows certain concentration dependent (such as Fig. 6
It is shown).
When 12.5 μM of Artesunate, 2.5 μM of Sorafenib, cell inhibitory rate reaches 50% or more, and experiment shows sweet wormwood
Amber ester can effectively combine the growth that Sorafenib inhibits liver cancer cells, next test will select 12.5 μM of Artesunate with
2.5 μM of Sorafenib combinations.
(2) cell clonal formation is tested
When cell grows into logarithmic phase, after being digested with pancreatin, the quantity of cell is measured with cell counter, is prepared clean
6 orifice plates, every hole number of plated cells are 5000.After culture for 24 hours, corresponding drug, respectively 12.5 μM of Artesunate, Suo La is added
2.5 μM of non-Buddhist nun, Artesunate combine (12.5 μM+2.5 μM) with Sorafenib, cultivate one in 37 DEG C, the incubator of 5%CO2
All (7 days) take out and use violet staining, and every hole is added 500 μ L Crystal Violet Dyes, is placed in 30 minutes on shaking table and is dyed, and use
PBS is cleaned 3 times, five minutes every time, is dried, is taken pictures after observation, and 10% glacial acetic acid of 1mL is added in every hole, is placed on shaking table 30 minutes
It decolourizes, every hole takes 100 μ L to be placed in 96 orifice plates, and microplate reader measures the absorbance under 550nm.
Data statistics: Cell colonies assay=ADosing group/AControl group；
Cell clone is as the result is shown: cell culture one week, see Fig. 7 and Fig. 8 as it can be seen that with blank control group, sweet wormwood amber is applied alone
Ester and Sorafenib group is applied alone to compare, Artesunate acts on human liver cancer cell Huh7 after combining with Sorafenib is significantly pressed down
System.
Effete test embodiment 4
Western blot detection Artesunate combines the variation to AKT/c-Myc protein expression with Sorafenib.
Huh7 cell is washed 3 times with the PBS of ice after the Sorafenib of various concentration and Artesunate processing, is added 200
The RIPA cell pyrolysis liquid (containing protease inhibitors) of μ l is collected into 1.5mlEP pipe, is inserted with cell scraper by under cell scraper
Enter and crack 1h in ice, during which overturn makes cell cracking abundant several times.Cell after cracking is centrifuged (12000rpm, 20min, 4 DEG C)
After take supernatant to measure its protein concentration with BCA, mixed with sample-loading buffer, 97 DEG C are boiled 10min, make albuminous degeneration.
20 μ l, SDS- polyacrylamide constant pressure 100v electrophoretic separation albumen is sampled, then extremely by the protein delivery on gel
On nitrocellulose filter, 2h is closed with the TBST containing 5% skimmed milk power, is washed 4 times, each 10min, 4 DEG C of primary antibody overnight incubations,
TBST is added secondary antibody (1:3000-1:5000) and is incubated at room temperature 60min after washing 4 times, TBST is washed 4 times, and ECL luminescent solution is added and exposes
Light.Developer solution 1:1 matching while using, drop covers film, darkroom tabletting, automatic film developer development, fixing on film when use.
Albumen is as the result is shown: to Huh7 cell independent role and combination, respectively Sorafenib (2.5 μM), Artesunate
After (12.5 μM) and Artesunate and Sorafenib combined treatment 48h, by western blot development detection (such as Fig. 9 and
Shown in Figure 10).
By Fig. 9 and Figure 10 the results show that compared with exclusive use, the table of p-AKT and c-Myc after two kinds of Drug combinations
It is significantly reduced up to amount.
The above described is only a preferred embodiment of the present invention, be not intended to limit the present invention in any form, therefore
Without departing from the technical solutions of the present invention, according to the technical essence of the invention it is to the above embodiments it is any modification,
Equivalent variations and modification, all of which are still within the scope of the technical scheme of the invention.
Claims (6)
1. Artesunate combines the application in the liver cancer Huh7 cell drug for preparing AKTc-Myc activation, feature with Sorafenib
Be: the Artesunate reduces the protein level of p-AKT, c-Myc, inhibits liver cancer Huh7 cell Proliferation, the Artesunate
The protein level by reducing p-AKT, c-Myc is combined with Sorafenib, further suppresses the growth of liver cancer Huh7 cell.
2. application according to claim 1, it is characterised in that: the Artesunate is from compositae plant artemisia annua
It is extracted in Artemisia annual, molecular formula C19H28O8, it is -10 α of dihydroartemisinine-succinate monoester, based on anhydride
It calculates, contains C19H28O8It should be 98.0%~102.0%, belong to white crystalline powder, odorless bitter, in ethyl alcohol, acetone or dichloro
Readily soluble in methane, adding methylene chloride makes to dissolve and quantify the solution for diluting and being made in every 1ml containing 25mg, measures in accordance with the law, specific rotation
It is+4.5 ° to+6.5 °, pH value is 3.5~4.5, and preparing fusing point is 1290 DEG C~1400 DEG C.
3. application according to claim 1, it is characterised in that: the Sorafenib is also known as Raf inhibitor, for white to ash
Color solid, molecular formula C21H16ClF3N4O3, it is slightly soluble in ethyl alcohol, is dissolved in polyethylene glycols 400, preparing fusing point is 202 DEG C~204
℃。
4. application according to claim 1, it is characterised in that: the Artesunate is combined with Sorafenib, Artesunate
Concentration >=12.5 μm olL-1, the concentration >=2.5 μm olL of Sorafenib-1。
5. application according to claim 1-3, it is characterised in that: the composition of medicine can be made into tablet, system
Preparation Method are as follows:
1) Artesunate 12.5g, Sorafenib 2.5g are weighed, starch 8.0g, dextrin 12.0g are added in mortar and are ground；
2) tartaric acid of 0.4g is dissolved in 50% ethyl alcohol, is repeatedly added in mixed-powder by appropriate amount, dispersion face is wanted when addition
Greatly, it is uniformly mixed, softwood is made；
3) wet grain is made by 18~20 mesh nylon mesh, 60 DEG C or less dryings can rise to 70 DEG C hereinafter, accelerate drying when close dry,
Dry granular moisture control is below 1.5%；
4) go out whole grain by 20 meshes, mix with the magnesium stearate of 0.4g sieving, then mixed again with dry particl, calculate content,
Punch die tabletting is carried out with 8mm punch die；Up to Artesunate Sorafenib piece.
6. application according to any one of claim 1-3, it is characterised in that: the composition of medicine can be made into freeze-dried powder
Agent, preparation method are as follows:
1) his ring paste of swelling hydroxypropyl times: essence weighs 180g HYDROXYPROPYL BETA-CYCLODEXTRIN, is placed in 1L beaker, and 400ml note is added
It penetrates with water, magnetic agitation (revolving speed 500rpm) 3h in 50 DEG C of water-baths, swelling is spare；
2) it prepares inclusion compound and weighs 2.5g Sorafenib and 12.5g Artesunate, be placed in 2L beaker, the hydroxypropyl being swollen is added
Base betadex solution, 800rpm stirs 10min until dissolution；
3) it adjusts pH and 72g mannitol is added, continue stirring to after dissolving, with the pH value of 1mol/L hydrochloric acid conditioning solution to 10.1,
Solution is settled to 900ml with water for injection；
4) 900mg active carbon is added in depyrogenation, after 500rpm magnetic agitation adsorbs 30min, 0.22 μm of filtering with microporous membrane；
5) filling, freeze-drying rolls lid to get Artesunate Sorafenib freeze drying powder injection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910350401.2A CN110025618A (en) | 2019-04-28 | 2019-04-28 | Artesunate combines the application in the liver cancer Huh7 cell drug for preparing AKTc-Myc activation with Sorafenib |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910350401.2A CN110025618A (en) | 2019-04-28 | 2019-04-28 | Artesunate combines the application in the liver cancer Huh7 cell drug for preparing AKTc-Myc activation with Sorafenib |
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| CN201910350401.2A Pending CN110025618A (en) | 2019-04-28 | 2019-04-28 | Artesunate combines the application in the liver cancer Huh7 cell drug for preparing AKTc-Myc activation with Sorafenib |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109602743A (en) * | 2018-11-22 | 2019-04-12 | 重庆赛拜欧生物医药科技有限公司 | The composition of medicine of a kind of Sorafenib and artemisine compounds and its preparing the application in anticancer drug |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109602743A (en) * | 2018-11-22 | 2019-04-12 | 重庆赛拜欧生物医药科技有限公司 | The composition of medicine of a kind of Sorafenib and artemisine compounds and its preparing the application in anticancer drug |
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Application publication date: 20190719 |